HETEROARYL SUBSTITUTED THIENO[2,3-d]PYRIMIDINE AND THEIR USE AS ADENOSINE A2a RECEPTOR ANTAGONISTS

ABSTRACT

This invention relates to a novel thieno[2,3-d]pyrimidine, Z, and its therapeutic and prophylactic uses, wherein X, R 1  and R 2  are defined in the specification. Disorders treated and/or prevented include Parkinson&#39;s Disease.

CROSS-REFERENCE TO RELATED APPLICATIONS

The present application claims the benefits of the filing of U.S.Provisional Application No. 61/104,791 filed Oct. 13, 2008. The completedisclosures of the aforementioned related patent applications are herebyincorporated herein by reference for all purposes.

FIELD OF THE INVENTION

This invention relates to a novel arylindenopyrimidine and itstherapeutic and prophylactic uses. Disorders treated and/or preventedinclude neurodegenerative and movement disorders ameliorated byantagonizing Adenosine A2a receptors.

BACKGROUND OF THE INVENTION

Adenosine A2a Receptors Adenosine is a purine nucleotide produced by allmetabolically active cells within the body. Adenosine exerts its effectsvia four subtypes of cell surface receptors (A1, A2a, A2b and A3), whichbelong to the G protein coupled receptor superfamily (Stiles, G. L.Journal of Biological Chemistry, 1992, 267, 6451). A1 and A3 couple toinhibitory G protein, while A2a and A2b couple to stimulatory G protein.A2a receptors are mainly found in the brain, both in neurons and glialcells (highest level in the striatum and nucleus accumbens, moderate tohigh level in olfactory tubercle, hypothalamus, and hippocampus etc.regions) (Rosin, D. L.; Robeva, A.; Woodard, R. L.; Guyenet, P. G.;Linden, J. Journal of Comparative Neurology, 1998, 401, 163).

In peripheral tissues, A2a receptors are found in platelets,neutrophils, vascular smooth muscle and endothelium (Gessi, S.; Varani,K.; Merighi, S.; Ongini, E.; Bores, P. A. British Journal ofPharmacology, 2000, 129, 2). The striatum is the main brain region forthe regulation of motor activity, particularly through its innervationfrom dopaminergic neurons originating in the substantial nigra. Thestriatum is the major target of the dopaminergic neuron degeneration inpatients with Parkinson's Disease (PD). Within the striatum, A2areceptors are co-localized with dopamine D2 receptors, suggesting animportant site for the integration of adenosine and dopamine signalingin the brain (Fink, J. S.; Weaver, D. Ri; Rivkees, S. A.; Peterfreund,R. A.; Pollack, A. E.; Adler, E. M.; Reppert, S. M. Brain ResearchMolecular Brain Research, 1992,14,186).

Neurochemical studies have shown that activation of A2a receptorsreduces the binding affinity of D2 agonist to their receptors. This D2Rand A2aR receptor-receptorinteraction has been demonstrated instriatalmembrane preparations of rats (Ferre, S.; con Euler, G.; Johansson, B.;Fredholm, B. B.; Fuxe, K. Proceedings of the National Academy ofSciences I of the United States of America, 1991, 88, 7238) as well asin fibroblast cell lines after transfected with A2aR and D2R cDNAs(Salim, H.; Ferre, S.; Dalal, A.; Peterfreund, R. A.; Fuxe, K.; Vincent,J. D.; Lledo, P. M. Journal of Neurochemistry, 2000, 74, 432). In vivo,pharmacological blockade of A2a receptors using A2a antagonist leads tobeneficial effects in dopaminergic neurotoxinMPTP(1-methyl-4-pheny-1,2,3,6-tetrahydropyridine)-induced PC) in variousspecies, including mice, rats, and monkeys (Ikeda, K.; Kurokawa, M.;Aoyana, S.; Kuwana, Y. Journal of Neurochemistry, 2002, 80, 262).

Furthermore, A2a knockout mice with genetic blockade of A2a functionhave been found to be less sensitive to motor impairment andneurochemical changes when they were exposed to neurotoxin MPTP (Chen,J. F.; Xu, K.; I Petzer, J. P.; Steal, R.; Xu, Y. H.; Beilstein, M.;Sonsalla, P. K.; Castagnoli, K.; Castagnoli, N., Jr.; Schwarsschild, M.A. Journal of Neuroscience, 2001, 1 21, RC 1 43).

In humans, the adenosine receptor antagonist theophylline has been foundto produce beneficial effects in PD patients (Mally, J.; Stone, T. W.Journal of the Neurological Sciences, 1995, 132, 129). Consistently,recent epidemiological study has shown that high caffeine consumptionmakes people less likely to develop PD (Ascherio, A.; Zhang, S. M.;Hernan, M. A.; Kawachi, I.; Colditz, G. A.; Speizer, F. E.; Willett, W.C. Annals of Neurology, 2001, 50, 56). In summary, adenosine A2areceptor blockers may provide a new class of antiparkinsonian agents(Impagnatiello, F.; Bastia, E.; Ongini, E.; Monopoli, A. EmergingTherapeutic Targets, 2000, 4, 635).

Antagonists of the A_(2A) receptor are potentially useful therapies forthe treatment of addiction. Major drugs of abuse (opiates, cocaine,ethanol, and the like) either directly or indirectly modulate dopaminesignaling in neurons particularly those found in the nucleus accumbens,which contain high levels of A_(2A) adenosine receptors. Dependence hasbeen shown to be augmented by the adenosine signaling pathway, and ithas been shown that administration of an A_(2A) receptor antagonistreduces the craving for addictive substances (“The Critical Role ofAdenosine A_(2A) Receptors and Gi βγ Subunits in Alcoholism andAddiction: From Cell Biology to Behavior”, by Ivan Diamond and Lina Yao,(The Cell Biology of Addiction, 2006, pp 291-316) and “Adaptations inAdenosine Signaling in Drug Dependence: Therapeutic Implications”, byStephen P. Hack and Macdonald J. Christie, Critical Review inNeurobiology, Vol. 15, 235-274 (2003)). See also Alcoholism: Clinicaland Experimental Research (2007), 31(8), 1302-1307.

An A_(2A) receptor antagonist could be used to treat attention deficithyperactivity disorder (ADHD) since caffeine (a non selective adenosineantagonist) can be useful for treating ADHD, and there are manyinteractions between dopamine and adenosine neurons. Clinical Genetics(2000), 58(1), 31-40 and references therein.

Antagonists of the A_(2A) receptor are potentially useful therapies forthe treatment of depression. A_(2A) antagonists are known to induceactivity in various models of depression including the forced swim andtail suspension tests. The positive response is mediated by dopaminergictransmission and is caused by a prolongation of escape-directed behaviorrather than by a motor stimulant effect. Neurology (2003), 61(suppl 6)S82-S87.

Antagonists of the A_(2A) receptor are potentially useful therapies forthe treatment of anxiety. A_(2A) antagonist have been shown to preventemotional/anxious responses in vivo. Neurobiology of Disease (2007),28(2) 197-205.

SUMMARY OF THE INVENTION

Compounds of Formula Z are potent small molecule antagonists of theAdenosine A2a receptor.

wherein

X is selected from the group consisting of

R¹ is heteroaryl which may be substituted with one substituent selectedfrom the group consisting of Cl, Br, F, OH, CN, C₍₁₋₄₎alkyl, CHF₂, CF₃,OCF₃, cyclopropyl, and OC₍₁₋₄₎alkyl;

R² is heteroaryl wherein said heteroaryl is optionally substituted withCl, F, Br, OC₍₁₋₄₎alkyl, OCF₃, OH, C₍₁₋₄₎alkyl, CHF₂, CF₃, OCH₂CF₃, or aring selected from the group consisting of:

-   -   wherein R^(a), R^(b), and R^(c) are independently H or        C₍₁₋₄₎alkyl;    -   R^(d) is H, —C₍₁₋₄₎alkyl, —CH₂CH₂OCH₂CH₂OCH₃, —CH₂CO₂H,        —C(O)C₍₁₋₄₎alkyl, or —CH₂C(O)C₍₁₋₄₎alkyl;

and solvates, hydrates, tautomers and pharmaceutically acceptable saltsthereof.

DETAILED DESCRIPTION OF THE INVENTION

Compounds of Formula Z are potent small molecule antagonists of theAdenosine A2a receptor.

wherein

X is selected from the group consisting of

R¹ is heteroaryl which may be substituted with one substituent selectedfrom the group consisting of Cl, Br, F, OH, CN, C₍₁₋₄₎alkyl, CHF₂, CF₃,OCF₃, cyclopropyl, and OC₍₁₋₄₎alkyl;

R² is heteroaryl wherein said heteroaryl is optionally substituted withCl, F, Br, OC₍₁₋₄₎alkyl, OCF₃, OH, C₁₋₄₎alkyl, CHF₂, CF₃, OCH₂CF₃, or aring selected from the group consisting of:

-   -   wherein R^(a), R^(b), and R^(c) are independently H or        C₍₁₋₄₎alkyl;    -   R^(d) is H, —C₍₁₋₄₎alkyl, —CH₂CH₂OCH₂CH₂OCH₃, —CH₂CO₂H,        —C(O)C₍₁₋₄₎alkyl, or —CH₂C(O)C₍₁₋₄₎alkyl;

and solvates, hydrates, tautomers and pharmaceutically acceptable saltsthereof.

In another embodiment of the invention:

X is selected from the group consisting of

R¹ is furyl, oxazolyl, thiazolyl, pyridyl, pyrimadyl, isoxazolyl,pyrrolyl, imidazoyl, or pyridazyl, any of which may be substituted withone substituent selected from the group consisting of Cl, Br, F, OH, CN,C₍₁₋₄₎alkyl, CHF₂, CF₃, cyclopropyl, and OC₍₁₋₄₎alkyl;

R² is pyrimadyl, isoxazolyl, pyrrolyl, imidazoyl, furyl, oxazolyl,pyridyl or pyridazyl, any of which may be substituted with onesubstituent selected from the group consisting of Cl, Br, F, OH,C₍₁₋₄₎alkyl, CHF₂, CF₃, and OC₍₁₋₄₎alkyl;

and solvates, hydrates, tautomers and pharmaceutically acceptable saltsthereof.

In another embodiment of the invention:

X is selected from the group consisting of

R¹ is furyl, oxazolyl, thiazolyl, pyridyl, or pyridazyl, any of whichmay be substituted with one substituent selected from the groupconsisting of Cl, Br, F, OH, CN, C₍₁₋₄₎alkyl, CHF₂, CF₃, cyclopropyl,and OC₍₁₋₄₎alkyl;

R² is pyridyl or pyridazyl, either of which may be substituted with onesubstituent selected from the group consisting of Cl, Br, F, OH,C₍₁₋₄₎alkyl, CHF₂, CF₃, and OC₍₁₋₄₎alkyl;

and solvates, hydrates, tautomers and pharmaceutically acceptable saltsthereof.

In another embodiment of the invention:

X is selected from the group consisting of

R¹ is furyl, oxazolyl, thiazolyl, pyridyl, or pyridazyl, any of whichmay be substituted with one substituent selected from the groupconsisting of CN, CH₃, CHF₂, cyclopropyl, and OCH₃;

R² is pyridyl or pyridazyl, either of which may be substituted with onesubstituent selected from the group consisting of Cl, Br, F, OH, andOCH₃;

and solvates, hydrates, tautomers and pharmaceutically acceptable saltsthereof.

In another embodiment of the invention:

X is selected from the group consisting of

R¹ is selected from the group consisting of:

R² is selected from the group consisting of:

and solvates, hydrates, tautomers and pharmaceutically acceptable saltsthereof.

Another embodiment of the invention is a compound selected from thegroup consisting of:

and solvates, hydrates, tautomers, and pharmaceutically acceptable saltsthereof.

This invention further provides a method of treating a subject having acondition ameliorated by antagonizing Adenosine A2a receptors, whichcomprises administering to the subject a therapeutically effective doseof a compound of Formula Z.

This invention further provides a method of preventing a disorderameliorated by antagonizing Adenosine A2a receptors in a subject,comprising of administering to the subject a prophylactically effectivedose of the compound of claim 1 either preceding or subsequent to anevent anticipated to cause a disorder ameliorated by antagonizingAdenosine A2a receptors in the subject.

Compounds of Formula Z can be isolated and used as free bases. They canalso be isolated and used as pharmaceutically acceptable salts.

Examples of such salts include hydrobromic, hydroiodic, hydrochloric,perchloric, sulfuric, maleic, fumaric, malic, tartaric, citric, adipic,benzoic, mandelic, methanesulfonic, hydroethanesulfonic,benzenesulfonic, oxalic, palmoic, 2 naphthalenesulfonic,p-toluenesulfonic, cyclohexanesulfamic and saccharic.

This invention also provides a pharmaceutical composition comprising acompound of Formula Z and a pharmaceutically acceptable carrier.

Pharmaceutically acceptable carriers are well known to those skilled inthe art and include, but are not limited to, from about 0.01 to about0.1 M and preferably 0.05 M phosphate buyer or 0.8% saline. Suchpharmaceutically acceptable carriers can be aqueous or non-aqueoussolutions, suspensions and emulsions. Examples of non-aqueous solventsare propylene glycol, polyethylene glycol, vegetable oils such as oliveoil, and injectable organic esters such as ethyl oleate. Aqueouscarriers include water, ethanol, alcoholic/aqueous solutions, glycerol,emulsions or suspensions, including saline and buffered media. Oralcarriers can be elixirs, syrups, capsules, tablets and the like. Thetypical solid carrier is an inert substance such as lactose, starch,glucose, methyl-cellulose, magnesium stearate, dicalcium phosphate,mannitol and the like. Parenteral carriers include sodium chloridesolution, Ringer's dextrose, dextrose and sodium chloride, lactatedRinger's and fixed oils. Intravenous carriers include fluid and nutrientreplenishers, electrolyte replenishers such as those based on Ringer'sdextrose and the like.

Preservatives and other additives can also be present, such as, forexample, antimicrobials, antioxidants, chelating agents, inert gases andthe like. All carriers can be mixed as needed with disintegrants,diluents, granulating agents, lubricants, binders and the like usingconventional techniques known in the art.

This invention further provides a method of treating a subject having acondition ameliorated by antagonizing Adenosine A2a receptors, whichcomprises administering to the subject a therapeutically effective doseof a compound of Formula Z.

In one embodiment, the disorder is a neurodegenerative or movementdisorder. Examples of disorders treatable by the instant pharmaceuticalcomposition include, without limitation, Parkinson's Disease,Huntington's Disease, Multiple System Atrophy, CorticobasalDegeneration, Alzheimer's Disease, and Senile Dementia.

In one preferred embodiment, the disorder is Parkinson's disease.

As used herein, the term “subject” includes, without limitation, anyanimal or artificially modified animal having a disorder ameliorated byantagonizing adenosine A2a receptors. In a preferred embodiment, thesubject is a human.

Administering the instant pharmaceutical composition can be effected orperformed using any of the various methods known to those skilled in theart. Compounds of Formula Z can be administered, for example,intravenously, intramuscularly, orally and subcutaneously. In thepreferred embodiment, the instant pharmaceutical composition isadministered orally. Additionally, administration can comprise givingthe subject a plurality of dosages over a suitable period of time. Suchadministration regimens can be determined according to routine methods.

As used herein, a “therapeutically effective dose” of a pharmaceuticalcomposition is an amount sufficient to stop, reverse or reduce theprogression of a disorder. A “prophylactically effective dose” of apharmaceutical composition is an amount sufficient to prevent adisorder, i.e., eliminate, ameliorate and/or delay the disorder's onset.Methods are known in the art for determining therapeutically andprophylactically effective doses for the instant pharmaceuticalcomposition. The effective dose for administering the pharmaceuticalcomposition to a human, for example, can be determined mathematicallyfrom the results of animal studies.

In one embodiment, the therapeutically and/or prophylactically effectivedose is a dose sufficient to deliver from about 0.001 mg/kg of bodyweight to about 200 mg/kg of body weight of a compound of Formula Z. Inanother embodiment, the therapeutically and/or prophylacticallyeffective dose is a dose sufficient to deliver from about 0.05 mg/kg ofbody weight to about 50 mg/kg of body weight. More specifically, in oneembodiment, oral doses range from about 0.05 mg/kg to about 100 mg/kgdaily. In another embodiment, oral doses range from about 0.05 mg/kg toabout 50 mg/kg daily, and in a further embodiment, from about 0.05 mg/kgto about 20 mg/kg daily. In yet another embodiment, infusion doses rangefrom about 1.0,ug/kg/min to about 10 mg/kg/min of inhibitor, admixedwith a pharmaceutical carrier over a period ranging from about severalminutes to about several days. In a further embodiment, for topicaladministration, the instant compound can be combined with apharmaceutical carrier at a drug/carrier ratio of from about 0.001 toabout 0.1.

The invention also provides a method of treating addiction in a mammal,comprising administering a therapeutically effective dose of a compoundof Formula Z.

The invention also provides a method of treating ADHD in a mammal,comprising administering a therapeutically effective dose of a compoundof Formula Z.

The invention also provides a method of treating depression in a mammal,comprising administering a therapeutically effective dose of a compoundof Formula Z.

The invention also provides a method of treating anxiety in a mammal,comprising administering a therapeutically effective dose of a compoundof Formula Z.

Definitions:

The term “C_(a-b)” (where a and b are integers referring to a designatednumber of carbon atoms) refers to an alkyl, alkenyl, alkynyl, alkoxy orcycloalkyl radical or to the alkyl portion of a radical in which alkylappears as the prefix root containing from a to b carbon atomsinclusive. For example, C₁₋₄ denotes a radical containing 1, 2, 3 or 4carbon atoms.

The term “alkyl,” whether used alone or as part of a substituent group,refers to a saturated branched or straight chain monovalent hydrocarbonradical, wherein the radical is derived by the removal of one hydrogenatom from a single carbon atom. Unless specifically indicated (e.g. bythe use of a limiting term such as “terminal carbon atom”), substituentvariables may be placed on any carbon chain atom. Typical alkyl radicalsinclude, but are not limited to, methyl, ethyl, propyl, isopropyl andthe like. Examples include C₁₋₈alkyl, C₁₋₆alkyl and C₁₋₄alkyl groups.

The term “heteroaryl” refers to a radical derived by the removal of onehydrogen atom from a ring carbon atom of a heteroaromatic ring system.Typical heteroaryl radicals include furyl, thienyl, pyrrolyl, oxazolyl,thiazolyl, imidazolyl, pyrazolyl, isoxazolyl, isothiazolyl, oxadiazolyl,triazolyl, thiadiazolyl, pyridinyl, pyridazinyl, pyrimidinyl, pyrazinyl,indolizinyl, indolyl, isoindolyl, benzo[b]furyl, benzo[b]thienyl,indazolyl, benzimidazolyl, benzthiazolyl, purinyl, 4H-quinolizinyl,quinolinyl, isoquinolinyl, cinnolinyl, phthalzinyl, quinazolinyl,quinoxalinyl, 1,8-naphthyridinyl, pteridinyl and the like.

Abbreviations:

Herein and throughout this application, the following abbreviations maybe used.

-   -   Cy cyclohexyl    -   DMF dimethylformamide    -   DMSO dimethylsulfoxide    -   Et ethyl    -   EtOAc ethyl acetate    -   KOtBu potassium tert-butoxide    -   Me methyl    -   NBS N-bromo succinimide    -   OAc acetate    -   Pd(dppf)Cl₂        [1,1′-Bis(diphenylphosphino)ferrocene]dichloropalladium (II)    -   py pyridine    -   THF tetrahydrofuran    -   Xantphos 9,9-Dimethyl-4,5-bis(diphenylphosphino)xanthene

General Schemes:

Compounds of Formula Z can be prepared by methods known to those who areskilled in the art. The following reaction schemes are only meant torepresent examples of the invention and are in no way meant to be alimit of the invention.

Scheme 1 illustrates the synthetic routes (Paths 1, 2 and 3) leading tocompounds of Formula Z (A, B, C, and D). Starting with2-amino-3-cyanothiophene I and following the path indicated by thearrows, condensation under basic conditions with R¹—CN, where R¹ is asdefined in Formula Z, affords the aminopyrimidine II. Theaminopyrimidine II is reacted with N-bromosuccinimide (NBS), to give thebromothiophene III. Following path 1 bromothiophene III is reacted withR²CH₂ZnCl or R²CH₂ZnBr, where R² is as defined in Formula Z, in thepresence of a palladium catalyst to afford compounds of Formula Z, whereX is CH₂ (A). Following path 2 bromothiophene III is reacted withdi-tert-butyldicarbonate [(Boc)₂O] in the presence of 4-dimethylaminopyridine (DMAP) to give IV that undergoes a metal-halogen exchange andis reacted with R²CHO, where R² is as defined in Formula Z to givecompounds V that are deprotected to give compounds where X is

Following path 3, alcohol V is oxidized with Dess-Martin periodinane togive VI that can be deprotected to give compounds of Formula Z, where X

Ketones VI are reacted with methylmagnesium chloride, followed by TFAmediated BOC removal, to afford compounds of Formula Z, where X is

Scheme 2 illustrates the synthetic routes (Paths 1, 2 and 3) leading tocompounds of Formula Z (A and B). Starting with 2-amino-3cyanothiopheneI and following the path indicated by the arrows, condensation underbasic conditions with R¹—CN, where R¹ is as defined in Formula Z,affords the aminopyrimidine II. The aminopyrimidine II is reacted withdi-tert-butyldicarbonate [(Boc)₂O] in the presence of 4-dimethylaminopyridine (DMAP) to give the corresponding protected amine IV. Thethiophene IV is deprotonated with lithium diisopropylamide (LDA) andreacted with R²CHO, where R² is as defined in Formula Z, to give anintermediate alcohol IX. Following path 1, IX is deprotected with TFA togive compounds of Formula B. Following path 2, IX is reacted withtriethylsilane in TFA to give compounds of Formula A. Following path 3,IX is reacted with acetic anhydride to give the corresponding acetate Xthat is reduced with samarium iodide (SmI₂) followed by TFA deprotectionto gives compounds of Formula A.

Scheme 3 illustrates the synthetic routes (Paths 1 and 2) leading tocompounds of Formula A and alkyl substituted compounds of Formula A.Starting with aldehyde XI, where R² is as defined in Formula Z, reactionwith malononitrile and elemental sulfur under basic conditions gives thethiophene XII. The thiophene XII is condensed under basic conditionswith R¹—CN, where R¹ is as defined in Formula Z, to afford compounds ofFormula Z where X is CR^(a)R^(b) and R^(a) and R^(b) are independentlyH, Me, or Et. Alternatively, aldehydes that are not commerciallyavailable can be synthesized following path 2 using XIII where R² is asdefined in Formula Z, is reacted with allyl bromide under basicconditions to give XIV. Alkene XIV is dihydroxylated using osmiumtetroxide in the presence of NMO to give diol XV. Oxidative cleavage ofXV with sodium periodate affords aldehydes XI that can follow the arrowsfrom path 1 to afford compounds of Formula A and alkyl substitutedcompounds of Formula A.

Scheme 4 illustrates the synthetic route to compounds of Formula R¹—CN,where R¹ is a C₍₁₋₄₎alkyl substituted furan. Scheme 4 also illustrateshow any R¹—CO₂CH₃ may be converted into R¹—CN. Bromofuran XVI can reactwith alkylzinc reagents in the presence of a palladium catalyst to giveXVII. Ester XVII (or any R¹—CO₂CH₃) is reacted with ammonium hydroxideto give the corresponding amide XVIII. Dehydration of the amide isaccomplished using POCl₃ in pyridine to give the desired heterocyclicnitrile R¹—CN.

Scheme 5 illustrates the synthetic routes (Paths 1, 2 and 3) leading tocompounds of Formula Z (E, F, G, and H). Following path 1,bromothiophene III is reacted with R²CH₂CH₂ZnCl or R²CH₂CH₂ZnBr, whereR² is as defined in Formula Z, in the presence of a palladium catalystto afford compounds of Formula Z, where X is CH₂CH₂ (E). Alternatively,compounds of Formula F can be reduced by hydrogenation to give compoundsof Formula Z, where X is

Following path 2 bromothiophene III is reacted with R²CHCHB(OH)₂, whereR² is as defined in Formula Z, in the presence of a palladium catalystto give compounds of Formula Z, where X is

Following path 3 bromothiophene III is reacted with R²C(CH₂)B(OH)₂,where R² is as defined in Formula Z, in the presence of palladium togive compounds of Formula Z where X is

Compounds of Formula G are reacted with trimethylsufoxonium iodide underbasic conditions to afford compounds of Formula Z, where X is

EXAMPLES Example 16-(6-Chloro-pyridin-3-ylmethyl)-2-(5-methyl-furan-2-yl)-thieno[2,3-d]pyrimidin-4-ylamineExample 1 Step a2-(5-Methyl-furan-2-yl)-thieno[2,3-d]pyrimidin-4-ylamine

Solid t-BuOK (904 mg, 8.1 mmol) was added to a dioxane suspension (20mL) of 2-amino-thiophene-3-carbonitrile (5.0 g, 40.3 mmol) and5-methyl-furan-2-carbonitrile (4.5 g, 40.3 mmol) and the mixture wasimmersed into a 130° C. oil bath. After 10 min the flask was removedfrom the oil bath, diluted with THF, filtered and dry packed onto silicagel. Column chromatography gave 5.8 g of2-(5-methyl-furan-2-yl)-thieno[2,3-d]pyrimidin-4-ylamine.

Example 1 Step b6-Bromo-2-(5-methyl-furan-2-yl)-thieno[2,3-d]pyrimidin-4-ylamine

Solid NBS (4.7 g, 26.4 mmol) was added to a THF solution (100 mL) of2-(5-methyl-furan-2-yl)-thieno[2,3-d]pyrimidin-4-ylamine (5.8 g, 25.1mmol). After 2 h the mixture was diluted with EtOAc and washedconsecutively with saturated aqueous NaHCO₃, 1 M aqueous Na₂S₂O₃, andbrine. The organic layer was dried (Na₂SO₄) and dry packed onto silicagel. Column chromatography gave 6.3 g of6-bromo-2-(5-methyl-furan-2-yl)-thieno[2,3-d]pyrimidin-4-ylamine.

Example 1 Step c6-(6-Chloro-pyridin-3-ylmethyl)-2-(5-methyl-furan-2-yl)-thieno[2,3-d]pyrimidin-4-ylamine

A 0.5 M THF solution of (6-chloro-3-pyridyl)methylzinc chloride (2.1 mL,1.06 mmol) was added to a THF solution (3 mL) of6-bromo-2-(5-methyl-furan-2-yl)-thieno[2,3-d]pyrimidin-4-ylamine (110mg, 0.35 mmol) and Pd(dppf)Cl₂ (29 mg, 0.04 mmol) and the mixture washeated to reflux. After 3 h the mixture was diluted with EtOAc, washedwith water then brine, dried (Na₂SO₄), and dry packed onto silica gel.Column chromatography gave 43 mg of the title compound. ¹H NMR (Acetone,400 MHz): δ=7.31-7.37 (m, 4H), 7.22-7.31 (m, 1H), 7.20 (d, J=1.3 Hz,1H), 7.02 (d, J=3.0 Hz, 1H), 6.72 (br. s., 2H), 6.15-6.20 (m, 1H), 4.23(s, 2H), 2.36 ppm (s, 3H); MS m/e 322 (M+H).

Example 22-Oxazol-2-yl-6-pyrazin-2-ylmethyl-thieno[2,3-d]pyrimidin-4-ylamineExample 2 Step a 2-But-3-enyl-pyrazine

A 2.5 M hexanes solution of n-BuLi (18.0 mL, 45 mmol) was added to a−78° C. THF solution (60 mL) of t-BuOK (5.1 g, 45 mmol) anddiisopropylamine (6.3 mL, 45 mmol). After 5 min at −78° C. the yellowmixture was warmed to −40° C. Neat methylpyrazine (2.7 mL, 30 mmol) wasadded and the mixture rapidly turned dark red. After 30 min at −40° C.the mixture was cooled to −78° C. and neat allyl bromide (7.6 mL, 90mmol) was added. After 30 min at −78° C. water was added and the mixturewas partially concentrated to remove volatile organics. The resultingmixture was extracted with dichloromethane and the combined organicswere dried (Na₂SO₄), concentrated, and purified via columnchromatography to give 2.2 g of 2-but-3-enyl-pyrazine.

Example 2 Step b 4-Pyrazin-2-yl-butane-1,2-diol

Osmium tetroxide (2.5 wt. % solution in t-BuOH, 4.0 mL, 0.32 mmol) wasadded to a 0° C. t-BuOH (30 mL)/water (30 mL) of 2-but-3-enyl-pyrazine(2.1 g, 15.8 mmol) and N-methyl morpholine N-oxide (2.0 g, 17.4 mmol)and the mixture was allowed to warm to rt overnight. TLC analysisindicated a low level of conversion, so an additional 8 mL of OsO₄ wasadded and the reaction mixture was stirred for 1 d. Conversion improved,but was still incomplete by TLC analysis; 0.5 equiv N-methyl morpholineN-oxide (925 mg) and 1.0 equiv pyridine (1.28 mL) were added, and themixture was stirred for 2 h. A solution of 24 g Na₂SO₃ in 96 mL waterwas added, and the mixture was partially concentrated to remove volatileorganics. The remaining aqueous solution was saturated with sodiumchloride and was exhaustively extracted with ethyl acetate. The organicextracts were dried (Na₂SO₄), concentrated, and was purified by columnchromatography to give 1.7 g of the title compound.

Example 2 Step c 3-Pyrazin-2-yl-propionaldehyde

An aqueous solution of sodium periodate (0.65 M, 20 mL, 13 mmol, 1.3equiv) was added to a suspension of silica gel (20 g) in dichloromethane(160 mL). A CH₂Cl₂ solution (10 mL) of 4-pyrazin-2-yl-butane-1,2-diol(1.7 g, 10.1 mmol) was then added. After 2 h the resulting white slurrywas vacuum filtered and washed with CH₂Cl₂. The filtrate was dried(Na₂SO₄) and concentrated to give 1.1 g of the title compound that wasused without further purification.

Example 2 Step d 2-Amino-5-pyrazin-2-ylmethyl-thiophene-3-carbonitrile

Solid elemental sulfur (257 mg, 8.0 mmol) was added to a 0° C. DMFsolution (2 mL) of 3-pyrazin-2-yl-propionaldehyde (1.1 g, 8.0 mmol) andEt₃N (0.67 mL, 4.8 mmol). After 1 h, the solution was cooled to 0° C.and solid malononitrile (529 mg, 8.0 mmol) was added and stirredovernight. The mixture was partitioned between EtOAc and saturatedaqueous sodium chloride, and the aqueous phase was extracted with EtOAc.The combined organic extracts were dried (Na₂SO₄), concentrated, andpurified by column chromatography to give 555 mg of the title compound.¹H NMR (CHLOROFORM-d, 300 MHz): δ (ppm) 8.44-8.59 (m, 3H), 6.54 (s, 1H),4.73 (br. s., 2H), 4.12 (s, 2H)

Example 2 Step e Oxazole-2-carboxylic acid amide

Oxazole-2-carboxylic acid ethyl ester (1.6 g, 11.4 mmol) was suspendedin concentrated NH₄OH (32 mL) and stirred vigorously. After 26 h theprecipitate was collected by vacuum filtration, affording 1.1 g of thetitle compound that was used without further purification.

Example 2 Step f Oxazole-2-carbonitrile

Neat POCl₃ (1.12 mL, 12.3 mmol) was added to a pyridine solution (17 mL)of oxazole-2-carboxylic acid amide (982 mg, 8.8 mmol). After 4 h themixture was cooled to 0° C. and taken to pH 3 with concentrated aqueousHCl. The aqueous mixture was extracted with Et₂O and the combinedextracts were washed with water then brine, dried (Mg₂SO₄), concentratedand used without further purification to give 478 mg of5-cyclopropyl-furan-2-carbonitrile. The residue contained water, and wastherefore dissolved in CH₂Cl₂, dried (Na₂SO₄), and concentrated to give573 mg of the title compound that was used without further purification.

Example 2 Step g2-Oxazol-2-yl-6-pyrazin-2-ylmethyl-thieno[2,3-d]pyrimidin-4-ylamine

Solid t-BuOK (7 mg, 0.06 mmol) was added to a dioxane suspension (0.20mL) of 2-amino-5-pyrazin-2-ylmethyl-thiophene-3-carbonitrile (68 mg,0.32 mmol) and oxazole-2-carbonitrile (33 mg, 0.35 mmol) and the mixturewas heated by microwave irradiation (150° C., 10 min, 300 W). Thereaction mixture was diluted with dichloromethane and methanol, drypacked onto silica gel, and purified via column chromatography to give77 mg of the title compound. ¹H NMR (DMSO-d₆, 300 MHz): δ (ppm) 8.76 (s,1H), 8.65 (s, 1H), 8.59 (s, 1H), 8.26 (s, 1H), 7.75 (br s, 2H), 7.42 (s,1H), 7.39 (s, 1H), 4.47 (s, 2H); MS m/e 311 (M+H).

Example 32-Oxazol-2-yl-6-pyridin-2-ylmethyl-thieno[2,3-d]pyrimidin-4-ylaminehydrochloride Example 3 Step a 3-Pyridin-2-yl-propionaldehyde

A CH₂Cl₂ solution (9 mL) of DMSO (3.7 mL, 52.5 mmol) was added to a −78°C. CH₂Cl₂ solution (20 mL) of oxalyl chloride (2.3 mL, 26.2 mmol)).After 10 min at −78° C. a CH₂Cl₂ solution (20 mL) of 2-pyridinepropanol(3.00 g, 21.9 mmol) was added. After 15 min at −78° C. neattriethylamine (15.2 mL, 109.3 mmol) was added, the mixture was stirredfor 5 min at −78° C., then allowed to warm to room temperature andstirred for an additional 40 min. Water was added to the blacksuspension and the resulting mixture was extracted with CH₂Cl₂. Thecombined organic extracts were dried (Na₂SO₄), concentrated, andpurified by column chromatography to give 1.2 g of the title compound.

Example 3 Step b 2-Amino-5-pyridin-2-ylmethyl-thiophene-3-carbonitrile

Solid elemental sulfur (239 mg, 7.5 mmol) was added to a 0° C. DMFsolution (2 mL) of 3-pyridin-2-yl-propionaldehyde (1.2 g, 9.0 mmol) andEt₃N (0.62 mL, 4.5 mmol). After 50 min, the solution was cooled to 0° C.and solid malononitrile (493 mg, 7.5 mmol) was added and the mixturewarmed to rt. After 40 min the mixture was partitioned between EtOAc andwater, and the aqueous phase was extracted with EtOAc. The combinedorganic extracts were dried (Na₂SO₄), concentrated, and purified bycolumn chromatography to give 540 mg of the title compound. ¹H NMR (300MHz, CHLOROFORM-d) δ ppm 8.56 (d, J=4.1 Hz, 1H), 7.64 (td, J=7.6, 1.7Hz, 1H), 7.13-7.24 (m, 2H), 6.50 (s, 1H), 4.65 (br. s., 2H), 4.09 (s,2H).

Example 3 Step c2-Oxazol-2-yl-6-pyridin-2-ylmethyl-thieno[2,3-d]pyrimidin-4-ylaminehydrochloride

Solid t-BuOK (9 mg, 0.08 mmol) was added to a dioxane suspension (0.20mL) of 2-amino-5-pyridin-2-ylmethyl-thiophene-3-carbonitrile (87 mg,0.40 mmol) and oxazole-2-carbonitrile (46 mg, 0.49 mmol, prepared as anintermediate in Example 2) and the mixture was heated by microwaveirradiation (130° C., 10 min, 300 W). The reaction mixture was dilutedwith dichloromethane and methanol, dry packed onto silica gel, andpurified via column chromatography to give 107 mg of the title compound.The free base was dissolved in CH₂Cl₂ containing a minimal amount ofmethanol to achieve solution and the solution was added to 1 M HCl inEt₂O. The precipitated hydrochloride salt was collected by vacuumfiltration to give 112 mg of the title compound. ¹H NMR (300 MHz,DMSO-D6) δ ppm 8.89 (d, J=4.9 Hz, 1H), 8.53 (t, J=7.7 Hz, 1H), 8.31 (s,1H), 7.92-8.18 (m, 4H), 7.51 (s, 1H), 7.48 (s, 1H), 4.80 (s, 2H); MS m/e310 (M+H).

Example 46-(1-Methyl-1-pyridin-2-yl-ethyl)-2-oxazol-2-yl-thieno[2,3-d]pyrimidin-4-ylamineExample 4 Step a 2-(1,1-Dimethyl-but-3-enyl)-pyridine

A 2.5 M hexanes solution of n-BuLi (18.0 mL, 45 mmol) was added to a−78° C. THF solution (60 mL) of t-BuOK (5.1 g, 45 mmol) anddiisopropylamine (6.3 mL, 45 mmol). After 5 min at −78° C. the yellowmixture was warmed to −40° C. After 15 min, neat 2-isopropylpyridine(3.87 mL, 30 mmol) was added and the mixture rapidly turned dark red.After 30 min at −40° C. the mixture was cooled to −78° C. and neat allylbromide (7.6 mL, 90 mmol) was added. After 30 min at −78° C. water wasadded and the mixture was partially concentrated to remove volatileorganics. The resulting mixture was extracted with dichloromethane andthe combined organics were dried (Na₂SO₄), concentrated, and purifiedvia column chromatography to give 4.3 g of2-(1,1-dimethyl-but-3-enyl)-pyridine.

Example 4 Step b 4-Methyl-4-pyridin-2-yl-pentane-1,2-diol

Osmium tetroxide (2.5 wt. % solution in t-BuOH, 13.4 mL, 1.1 mmol) wasadded to a 0° C. t-BuOH (40 mL)/water (40 mL) of2-(1,1-dimethyl-but-3-enyl)-pyridine (3.5 g, 21.4 mmol) and N-methylmorpholine N-oxide (2.8 g, 23.6 mmol) and the mixture was allowed towarm to rt. After 3 h solid Na₂SO₃ (32 g) was added portionwise and theresulting suspension was stirred for 1 h. The mixture was partitionedbetween water and EtOAc and the aqueous phase was extracted with EtOAc.The combined organic extracts were dried (Na₂SO₄), concentrated, and waspurified by column chromatography to give 3.9 g of the title compound.

Example 4 Step c 3-Methyl-3-pyridin-2-yl-butyraldehyde

An aqueous solution of sodium periodate (0.65 M, 20 mL, 13 mmol) wasadded to a suspension of silica gel (20 g) in dichloromethane (160 mL).A CH₂Cl₂ solution (10 mL) solution of4-methyl-4-pyridin-2-yl-pentane-1,2-diol (2.0 g, 10.0 mmol) was thenadded. After 1.5 h the resulting white slurry was vacuum filtered andwashed with CH₂Cl₂. The filtrate was dried (Na₂SO₄) and concentrated togive 682 mg of the title compound.

Example 4 Step d2-Amino-5-(1-methyl-1-pyridin-2-yl-ethyl)-thiophene-3-carbonitrile

Solid elemental sulfur (110 mg, 3.4 mmol) was added to a 0° C. DMFsolution (1 mL) of 3-methyl-3-pyridin-2-yl-butyraldehyde (671 mg, 4.1mmol) and Et₃N (0.29 mL, 2.1 mmol). After 50 min, the solution wascooled to 0° C. and solid malononitrile (226 mg, 3.4 mmol) was added andstirred overnight. The mixture was partitioned between EtOAc andsaturated aqueous sodium chloride, and the aqueous phase was extractedwith EtOAc. The combined organic extracts were dried (Na₂SO₄),concentrated, and purified by column chromatography to give 430 mg ofthe title compound. ¹H NMR (CHLOROFORM-d, 300 MHz): ¹H NMR(CHLOROFORM-d, 300 MHz): δ (ppm) 8.57 (d, J=4.9 Hz, 1H), 7.61 (td,J=7.8, 2.1 Hz, 1H), 7.25-7.28 (m, 1H, obscured by CHCl₃ peak), 7.24 (dt,J=7.9, 1.1, 1H), 7.14 (ddd, J=7.5, 4.9, 1.1 Hz, 1H), 6.49 (s, 1H), 4.63(br. s., 2H), 1.73 (s, 6H).

Example 4 Step e6-(1-Methyl-1-pyridin-2-yl-ethyl)-2-oxazol-2-yl-thieno[2,3-d]pyrimidin-4-ylamine

Solid t-BuOK (6 mg, 0.05 mmol) was added to a dioxane suspension (0.20mL) of2-amino-5-(1-methyl-1-pyridin-2-yl-ethyl)-thiophene-3-carbonitrile (63mg, 0.26 mmol) and oxazole-2-carbonitrile (27 mg, 0.28 mmol, prepared asan intermediate in Example 2) and the mixture was heated by microwaveirradiation (130° C., 10 min, 300 W). The reaction mixture was dilutedwith dichloromethane and methanol, dry packed onto silica gel, andpurified via column chromatography to give 44 mg of the title compound.¹H NMR (DMSO-d₆, 300 MHz): δ (ppm) 8.57 (ddd, J=4.7, 1.1, 0.9 Hz, 1H),8.25 (s, 1H), 7.69-7.80 (m, 3H), 7.53 (s, 1H), 7.42 (s, 1H), 7.40 (d,J=7.9 Hz, 1H), 7.24-7.29 (m, 1H), 1.83 (s, 6H); MS m/e 338 (M+H).

Example 56-(1-Methyl-1-pyridin-2-yl-ethyl)-2-pyrazin-2-yl-thieno[2,3-d]pyrimidin-4-ylamine

The title compound was prepared using pyrazinecarbonitrile in place ofoxazole-2-carbonitrile as described in Example 4. ¹H NMR (DMSO-d₆, 300MHz): δ (ppm) 9.45 (d, J=1.1 Hz, 1H), 8.72-8.75 (m, 1H), 8.70 (d, J=2.3Hz, 1H), 8.55-8.59 (m, 1H), 7.76 (td, J=7.7, 1.8 Hz, 1H), 7.68 (br s,2H), 7.54 (s, 1H), 7.40 (d, J=7.9 Hz, 1H), 7.27 (dd, J=7.4, 4.8 Hz, 1H),1.84 (s, 6H); MS m/e 349 (M+H).

Example 62-(5-Methyl-furan-2-yl)-6-pyrazin-2-ylmethyl-thieno[2,3-d]pyrimidin-4-ylamine

The title compound was prepared using 5-methyl-2-furonitrile in place ofoxazole-2-carbonitrile as described in Example 2. ¹H NMR (DMSO-d₆, 300MHz): δ (ppm) 8.74 (s, 1H), 8.63 (s, 1H), 8.58 (s, 1H), 7.48 (br s, 2H),7.29 (s, 1H), 7.00 (d, J=3.0 Hz, 1H), 6.25 (d, J=3.0 Hz, 1H), 4.41 (s,2H), 2.35 (s, 3H); MS m/e 324 (M+H).

Example 72-(4-Methyl-thiazol-2-yl)-6-pyrazin-2-ylmethyl-thieno[2,3-d]pyrimidin-4-ylamine

The title compound was prepared using 4-methylthiazole-2-carbonitrile inplace of oxazole-2-carbonitrile as described in Example 2. ¹H NMR(DMSO-d₆, 300 MHz): δ (ppm) 8.75 (s, 1H), 8.63-8.66 (m, 1H), 8.59 (d,J=2.6 Hz, 1H), 7.69 (br s, 2H), 7.40 (s, 1H), 7.37 (s, 1H), 4.45 (s,2H), 2.43 (s, 3H); MS m/e 341 (M+H).

Example 82-(5-Methyl-furan-2-yl)-6-(1-methyl-1-pyridin-2-yl-ethyl)-thieno[2,3-d]pyrimidin-4-ylamine

The title compound was prepared using 5-methyl-2-furonitrile in place ofoxazole-2-carbonitrile as described in Example 4. ¹H NMR (DMSO-d₆, 300MHz): δ (ppm) 8.56 (d, J=4.9 Hz, 1H), 7.74 (dt, J=7.7, 1.9 Hz, 1H), 7.47(br s, 2H), 7.43 (s, 1H), 7.36 (d, J=7.9 Hz, 1H), 7.25 (dd, J=7.5, 4.9Hz, 1H), 6.99 (d, J=3.0 Hz, 1H), 6.24 (d, J=3.0 Hz, 1H), 2.35 (s, 3H),1.80 (s, 6H); MS m/e 351 (M+H).

Example 92-Oxazol-5-yl-6-pyrazin-2-ylmethyl-thieno[2,3-d]pyrimidin-4-ylamine

The title compound was prepared using oxazole-5-carbonitrile in place ofoxazole-2-carbonitrile as described in Example 2. ¹H NMR (DMSO-d₆, 300MHz): δ (ppm) 8.74 (s, 1H), 8.64 (s, 1H), 8.58 (d, J=2.6 Hz, 1H), 8.51(s, 1H), 7.73 (s, 1H), 7.65 (br s, 2H), 7.34 (s, 1H), 4.44 (s, 2H); MSm/e 311 (M+H).

Example 10(±)-2-(5-Methyl-furan-2-yl)-6-(1-pyridin-2-yl-propyl)-thieno[2,3-d]pyrimidin-4-ylaminehydrochloride

The title compound was prepared using 2-n-propylpyridine and5-methyl-2-furonitrile in place of 2-isopropylpyridine andoxazole-2-carbonitrile, respectively as described in Example 4. ¹H NMR(DMSO-d₆, 300 MHz): δ (ppm) 8.77 (d, J=5.3 Hz, 1H), 8.25 (t, J=7.5 Hz,1H), 7.81 (d, J=7.9 Hz, 1H), 7.65-7.73 (m, 2H), 7.49 (d, J=3.0 Hz, 1H),6.42 (d, J=3.4 Hz, 1H), 4.68 (t, J=7.5 Hz, 1H), 2.41 (s, 3H), 2.14-2.31(m, 2H), 0.91 (t, J=7.2 Hz, 3H); MS m/e 351 (M+H).

Example 11(±)-2-(4-Methyl-thiazol-2-yl)-6-(1-pyridin-2-yl-propyl)-thieno[2,3-d]pyrimidin-4-ylaminehydrochloride

The title compound was prepared using 2-n-propylpyridine and4-methylthiazole-2-carbonitrile in place of 2-isopropylpyridine andoxazole-2-carbonitrile, respectively, as described in Example 4. ¹H NMR(DMSO-d₆, 300 MHz): δ (ppm) 8.75 (d, J=4.9 Hz, 1H), 8.20 (t, J=7.5 Hz,1H), 7.78 (d, J=7.5 Hz, 1H), 7.61-7.69 (m, 2H), 7.57 (s, 1H), 4.65 (t,J=7.7 Hz, 1H), 2.14-2.31 (m, 2H), 0.91 (t, J=7.2 Hz, 3H); MS m/e 368(M+H).

Example 12(±)-2-Oxazol-2-yl-6-(1-pyridin-2-yl-propyl)-thieno[2,3-d]pyrimidin-4-ylaminehydrochloride

The title compound was prepared using 2-n-propylpyridine in place of2-isopropylpyridine as described in Example 4. ¹H NMR (DMSO-d₆, 300MHz): δ (ppm) 8.81 (d, J=4.9 Hz, 1H), 8.29-8.40 (m, 2H), 7.90 (d, J=7.9Hz, 1H), 7.78 (t, J=6.4 Hz, 1H), 7.62 (s, 1H), 7.47 (s, 1H), 4.75 (t,J=7.5 Hz, 1H), 2.18-2.34 (m, 2H), 0.92 (t, J=7.3 Hz, 3H); MS m/e 338(M+H).

Example 136-(1-Methyl-1-pyridin-2-yl-ethyl)-2-(4-methyl-thiazol-2-yl)-thieno[2,3-d]pyrimidin-4-ylamine

The title compound was prepared using 4-methylthiazole-2-carbonitrile inplace of oxazole-2-carbonitrile as described in Example 4. ¹H NMR(DMSO-d₆, 300 MHz): δ (ppm) 8.57 (d, J=3.8 Hz, 1H), 7.76 (td, J=7.7, 1.9Hz, 1H), 7.67 (br s, 2H), 7.51 (s, 1H), 7.39 (t, J=4.0 Hz, 2H), 7.27(dd, J=7.0, 5.1 Hz, 1H), 2.43 (s, 3H), 1.83 (s, 6H); MS m/e 368 (M+H).

Example 14(±)-2-Oxazol-5-yl-6-(1-pyridin-2-yl-propyl)-thieno[2,3-d]pyrimidin-4-ylaminehydrochloride

The title compound was prepared using 2-n-propylpyridine andoxazole-5-carbonitrile in place of 2-isopropylpyridine andoxazole-2-carbonitrile, respectively, as described in Example 4. ¹H NMR(DMSO-d₆, 300 MHz): δ(ppm) 8.78 (d, J=4.9 Hz, 1H), 8.54 (s, 1H), 8.28(t, J=8.1 Hz, 1H), 7.67-7.90 (m, 4H), 7.53 (s, 1H), 4.65 (t, J=7.5 Hz,1H), 2.16-2.32 (m, 2H), 0.91 (t, J=7.3 Hz, 3H); MS m/e 338 (M+H).

Example 152-Oxazol-5-yl-6-pyridin-2-ylmethyl-thieno[2,3-d]pyrimidin-4-ylamine

The title compound was prepared using oxazole-5-carbonitrile in place ofoxazole-2-carbonitrile as described in Example 3. 1H NMR (300 MHz,CHLOROFORM-D) δ ppm 8.59 (d, J=4.1 Hz, 1H), 8.00 (s, 1H), 7.82 (s, 1H),7.67 (td, J=7.6, 1.7 Hz, 1H), 7.18-7.31 (m, 2H), 6.93 (s, 1H), 5.44 (brs, 2H), 4.37 (s, 2H); MS m/e 310 (M+H).

Example 162-(5-Methyl-furan-2-yl)-6-pyridin-2-ylmethyl-thieno[2,3-d]pyrimidin-4-ylamine

The title compound was prepared using 5-methyl-2-furonitrile in place ofoxazole-2-carbonitrile as described in Example 3. 1H NMR (300 MHz,CHLOROFORM-D) δ ppm 8.54 (d, J=3.8 Hz, 1H), 7.62 (td, J=7.6, 1.7 Hz,1H), 7.23 (d, J=7.9 Hz, 1H), 7.13-7.17 (m, 2H), 6.86 (s, 1H), 6.12 (d,J=2.3 Hz, 1H), 5.61 (s, 2H), 4.31 (s, 2H), 2.41 (s, 3H); MS m/e 322(M+H).

Example 172-(4-Methyl-thiazol-2-yl)-6-pyridin-2-ylmethyl-thieno[2,3-d]pyrimidin-4-ylamine

The title compound was prepared using 4-methylthiazole-2-carbonitrile inplace of oxazole-2-carbonitrile as described in Example 3. 1H NMR (300MHz, CHLOROFORM-D) δ ppm 8.59 (d, J=4.1 Hz, 1H), 7.67 (td, J=7.7, 1.9Hz, 1H), 7.17-7.29 (m, 2H), 7.02 (s, 1H), 6.93 (s, 1H), 5.46 (br s, 2H),4.37 (s, 2H), 2.56 (s, 3H); MS m/e 340 (M+H).

Example 182-Oxazol-2-yl-6-pyridin-3-ylmethyl-thieno[2,3-d]pyrimidin-4-ylamine

The title compound was prepared using 3-pyridinepropanol in place of2-pyridinepropanol as described in Example 3. 1H NMR (300 MHz, MeOD) δppm 8.55 (d, J=1.9 Hz, 1H), 8.46 (dd, J=4.7, 1.3 Hz, 1H), 8.09 (s, 1H),7.82 (d, J=7.9 Hz, 1H), 7.43 (dd, J=7.7, 5.1 Hz, 1H), 7.40 (s, 1H), 7.24(s, 1H), 4.33 (s, 2H); MS m/e 310 (M+H).

Example 192-(5-Cyclopropyl-furan-2-yl)-6-pyridin-3-ylmethyl-thieno[2,3-d]pyrimidin-4-ylamineExample 19 Step a 5-Cyclopropyl-furan-2-carboxylic acid methyl ester

Solid cyclopropylboronic acid (575 mg, 6.7 mmol) was added to a toluene(22 mL)/water (1.1 mL) solution of 5-bromo-furan-2-carboxylic acidmethyl ester (980 mg, 4.8 mmol), Pd(OAc)₂ (54 mg, 0.2 mmol), P(Cy)₃ (135mg, 0.5 mmol), and K₃PO₄ (3.6 g, 16.8 mmol). The resulting mixture washeated to 90° C. After 5 h the mixture was cooled, filtered andextracted with EtOAc. The combined organic extracts were washed withwater and brine, dried (Na₂SO₄), concentrated and purified via columnchromatography to give 650 mg of 5-cyclopropyl-furan-2-carboxylic acidmethyl ester.

Example 19 Step b 5-Cyclopropyl-furan-2-carboxylic acid amide

5-cyclopropyl-furan-2-carboxylic acid methyl ester (650 mg, 3.9 mmol)was suspended in concentrated NH₄OH (20 mL) and stirred vigorously.After 16 h the mixture was diluted with water and the aqueous phase wasextracted with EtOAc. The combined organic extracts were washed withwater and brine, dried (Na₂SO₄), concentrated and used without furtherpurification to give 550 mg of 5-cyclopropyl-furan-2-carboxylic acidamide.

Example 19 Step c 5-Cyclopropyl-furan-2-carbonitrile

Neat POCl₃ (0.48 mL, 5.1 mmol) was added to a pyridine solution (9 mL)of 5-cyclopropyl-furan-2-carboxylic acid amide (550 mg, 3.6 mmol). After2 h the mixture was cooled to 0° C. and taken to pH 4.5 withconcentrated aqueous HCl. The aqueous mixture was extracted with Et₂Oand the combined extracts were washed with brine, dried (Na₂SO₄),concentrated and used without further purification to give 478 mg of5-cyclopropyl-furan-2-carbonitrile.

Example 19 Step d2-(5-Cyclopropyl-furan-2-yl)-6-pyridin-3-ylmethyl-thieno[2,3-d]pyrimidin-4-ylamine

The title compound was prepared using 3-pyridinepropanol and5-cyclopropyl-furan-2-carbonitrile in place of 2-pyridinepropanol andoxazole-2-carbonitrile, respectively as described in Example 3. ¹H NMR(DMSO-d₆, 300 MHz): δ=8.57 (d, J=1.9 Hz, 1H), 8.49 (dd, J=4.9, 1.5 Hz,1H), 7.68-7.77 (m, 1H), 7.45 (s, 1H), 7.23 (s, 1H), 6.99 (d, J=3.4 Hz,1H), 6.20 (d, J=3.4 Hz, 1H), 4.24 (s, 2H), 3.33 (s, 2H), 2.01 (s, 1H),0.89-0.99 (m, 2H), 0.71-0.80 ppm (m, 2H); MS m/e 349 (M+H).

Example 206-Benzyl-2-(5-cyclopropyl-furan-2-yl)-thieno[2,3-d]pyrimidin-4-ylaminehydrochloride

The title compound was prepared using 5-cyclopropyl-furan-2-carbonitrilein place of and oxazole-2-carbonitrile as described in Example 3. ¹H NMR(DMSO-d₆, 300 MHz): δ=8.84 (br. s., 1H), 8.38 (br. s., 1H), 7.87 (d,J=8.3 Hz, 2H), 7.51 (s, 1H), 7.39-7.44 (m, 1H), 6.40 (d, J=3.0 Hz, 1H),4.69 (s, 2H), 2.08 (t, J=5.1 Hz, 1H), 1.24 (br. s., 2H), 0.86 ppm (s,2H); MS m/e 349 (M+H).

Example 216-(4-Amino-6-pyridin-3-ylmethyl-thieno[2,3-d]pyrimidin-2-yl)-pyridine-2-carbonitrile

The title compound was prepared using 3-pyridinepropanol andpyridine-2,6-dicarbonitrile in place of 2-pyridinepropanol andoxazole-2-carbonitrile, respectively as described in Example 3. ¹H NMR(CHLOROFORM-d, 300 MHz): δ=8.02 (d, J=6.8 Hz, 2H), 7.45-7.71 (m, 4H),7.19 (s, 1H), 6.99 (dd, J=8.7, 1.9 Hz, 1H), 3.83 (s, 2H), 3.3 ppm (s,2H); MS m/e 345 (M+H).

Example 22[4-Amino-2-(2-methoxy-pyridin-4-yl)-thieno[2,3-d]pyrimidin-6-yl]-phenyl-methanolExample 22 Step a2-(2-Methoxy-pyridin-4-yl)-thieno[2,3-d]pyrimidin-4-ylamine

The title compound was prepared using 2-methoxy-isonicotinonitrile andin place of 5-methyl-furan-2-carbonitrile as described in Example 1.

Example 22 Step b[2-(2-Methoxy-pyridin-4-yl)-thieno[2,3-d]pyrimidin-4-yl]-bis-carbamicacid tert-butyl ester

Solid DMAP (100 mg, 0.82 mmol) was added to a THF solution (20 mL) of2-(2-methoxy-pyridin-4-yl)-thieno[2,3-d]pyrimidin-4-ylamine (2.0 g, 8.0mmol) and (Boc)₂O (4.4 g, 20.2 mmol). After 2 h the mixture wasconcentrated in vacuo, and the resulting solid was diluted with CH₂Cl₂,filtered, and the filtrate was concentrated and purified by columnchromatography to give 3.0 g of the title compound.

Example 22 Step c[6-(Hydroxy-pyridin-2-yl-methyl)-2-(2-methoxy-pyridin-4-yl)-thieno[2,3-d]pyrimidin-4-yl]-bis-carbamicacid tert-butyl ester

A 1.8 M LDA solution (0.54 mL, 0.97 mmol) was added to a −78° C. THFsolution (3.5 mL) of[2-(2-methoxy-pyridin-4-yl)-thieno[2,3-d]pyrimidin-4-yl]-bis-carbamicacid tert-butyl ester (400 mg, 0.87 mmol). After 8 min, neatpyridine-2-carbaldehyde (140 mg, 1.31 mmol) was added and the mixturewas allowed to warm to −20° C. over 40 min. Saturated aqueous NH₄Cl wasadded and the layers were separated. The aqueous layer was extractedwith CH₂Cl₂ and the combined organics were dried (Na₂SO₄), concentrated,and purified by column chromatography to give 200 mg of the titlecompound.

Example 22 Step d[4-Amino-2-(2-methoxy-pyridin-4-yl)-thieno[2,3-d]pyrimidin-6-yl]-phenyl-methanol

Neat TFA (0.15 mL) was added to a CH₂Cl₂ solution (0.4 mL) of[6-(hydroxy-phenyl-methyl)-2-(2-methoxy-pyridin-4-yl)-thieno[2,3-d]pyrimidin-4-yl]-bis-carbamicacid tert-butyl ester (40 mg). After 1 h the mixture was concentratedand the resulting solid was partitioned between CH₂Cl₂ and saturatedaqueous NaHCO₃. The organic phase was separated, dried (Na₂SO₄), andconcentrated to provide 37 mg of the title compound. ¹H NMR (300 MHz,Acetone-d₆) δ=8.56 (d, J=4.9 Hz, 1H), 8.22 (d, J=5.3 Hz, 1H), 7.79-7.92(m, 2H), 7.62-7.72 (m, 2H), 7.49 (s, 1H), 7.32 (dd, J=4.9, 7.5 Hz, 1H),6.99 (br. s., 2H), 6.13 (d, J=4.5 Hz, 1H), 5.92 (br. s., 1H), 3.92 (s,3H); MS m/e 366 (M+H).

Example 23[4-Amino-2-(5-difluoromethyl-furan-2-yl)-thieno[2,3-d]pyrimidin-6-yl]-(3-fluoro-pyridin-2-yl)-methanolExample 23 Step a 5-Difluoromethyl-furan-2-carbonitrile

To a solution of Et₂NSF₃ (2.8 mL, 21.4 mmol) and CH₂Cl₂ (10 mL) at 4° C.was added a solution of 5-formyl-furan-2-carbonitrile (2.44 g, 20.2mmol; W. Hoyle and G. P. Roberts, J. Med. Chem. 1973, 16, 709) in CH₂Cl₂(10 mL). After 30 min at 4° C., saturated aqueous NaHCO₃ was added, thelayers were separated and the aqueous layer was extracted with CH₂Cl₂.The combined organics were dried (Na₂SO₄) and concentrated to give 2.15g of 5-difluoromethyl-furan-2-carbonitrile that was used without furtherpurification.

Example 23 Step b2-(5-Difluoromethyl-furan-2-yl)-thieno[2,3-d]pyrimidin-4-ylamine

Solid t-BuONa (280 mg, 2.9 mmol) was added to a dioxane solution (8 mL)of 5-difluoromethyl-furan-2-carbonitrile (2.1 g, 14.7 mmol) and2-amino-thiophene-3-carbonitrile (1.8 g, 14.7 mmol) and the mixtureimmediately became very hot, some solid was formed, and stirring becamedifficult. An additional 4 mL of dioxane was added and the mixture wasstirred. After 1.5 h the mixture was diluted with THF and dry packedonto silica gel. Column chromatography gave 2.9 g of the title compound.

Example 23 Step c6-Bromo-2-(5-difluoromethyl-furan-2-yl)-thieno[2,3-d]pyrimidin-4-ylamine

Solid NBS (3.0 g, 16.6 mmol) was added to a THF solution (110 mL) of2-(5-difluoromethyl-furan-2-yl)-thieno[2,3-d]pyrimidin-4-ylamine (3.1 g,11.8 mmol). After 2.5 h the mixture was dry packed onto silica gel.Column chromatography gave 4.8 g of the title compound.

Example 23 Step d[6-Bromo-2-(5-difluoromethyl-furan-2-yl)-thieno[2,3-d]pyrimidin-4-yl]-bis-carbamicacid tert-butyl ester

Solid DMAP (170 mg, 1.39 mmol) was added to a THF solution (130 mL) of6-bromo-2-(5-difluoromethyl-furan-2-yl)-thieno[2,3-d]pyrimidin-4-ylamine(4.8 g, 13.9 mmol) and Boc₂O (7.93 g, 36.4 mmol). After 40 min themixture was concentrated in vacuo and purified via column chromatographyto afford 4.6 g of the title compound.

Example 23 Step e{2-(5-Difluoromethyl-furan-2-yl)-6-[(3-fluoro-pyridin-2-yl)-hydroxy-methyl]-thieno[2,3-d]pyrimidin-4-yl}-bis-carbamicacid tert-butyl ester

A 1.0 M THF solution of i-PrMgCl.LiCl (0.70 mL, 0.70 mmol) was added toa −78° C. THF solution (1 mL) of[6-bromo-2-(5-difluoromethyl-furan-2-yl)-thieno[2,3-d]pyrimidin-4-yl]-bis-carbamicacid tert-butyl ester (308 mg, 0.56 mmol). After 1 h at −78° C. a THFsolution (0.8 mL) of 3-fluoro-pyridine-2-carbaldehyde (106 mg, 0.85mmol) was added and the reaction mixture was warmed to −15° C. After 40min, saturated aqueous NH₄Cl was added and the aqueous phase wasextracted with CH₂Cl₂. The combined organics were dried (Na₂SO₄),concentrated, and purified via column chromatography to give 183 mg ofthe title compound.

Example 23 Step f[4-Amino-2-(5-difluoromethyl-furan-2-yl)-thieno[2,3-d]pyrimidin-6-yl]-(3-fluoro-pyridin-2-yl)-methanol

Neat TFA (0.2 mL) was added to a CH₂Cl₂ solution (0.8 mL) of{2-(5-difluoromethyl-furan-2-yl)-6-[(3-fluoro-pyridin-2-yl)-hydroxy-methyl]-thieno[2,3-d]pyrimidin-4-yl}-bis-carbamicacid tert-butyl ester (42 mg, 0.07 mmol). After 1 h the mixture wasconcentrated in vacuo and the residue was partitioned between saturatedaqueous NaHCO₃ and EtOAc. The layers were separated and the organiclayer was dried (Na₂SO₄) and concentrated to give 16 mg of the titlecompound. ¹H NMR (300 MHz, Acetone-d₆) δ=8.49 (d, J=4.5 Hz, 1H),7.62-7.75 (m, 1H), 7.51 (dt, J=4.3, 8.4 Hz, 1H), 7.40 (s, 1H), 7.14-7.25(m, 1H), 7.00 (t, J=53.7 Hz, 1H), 6.89-7.09 (m, 3H), 6.33 (s, 1H), 5.63(br. s., 1H); MS m/e 393 (M+H).

Example 242-(5-Difluoromethyl-furan-2-yl)-6-(3-fluoro-pyridin-2-ylmethyl)-thieno[2,3-d]pyrimidin-4-ylamineExample 24 Step a Acetic acid[4-tert-butoxycarbonylamino-2-(5-difluoromethyl-furan-2-yl)-thieno[2,3-d]pyrimidin-6-yl]-(3-fluoro-pyridin-2-yl)-methylester

Solid DMAP (5 mg, 0.04 mmol) was added to a CH₂Cl₂ solution (1 mL) of{2-(5-difluoromethyl-furan-2-yl)-6-[(3-fluoro-pyridin-2-yl)-hydroxy-methyl]-thieno[2,3-d]pyrimidin-4-yl}-bis-carbamicacid tert-butyl ester (120 mg, 0.20 mmol, an intermediate prepared inExample 23), Ac₂O (50 μL, 0.53 mmol) and pyridine (0.1 mL). After 5 hthe mixture was concentrated and purified by flash chromatography togive 110 mg of the title compound.

Example 24 Step b[2-(5-Difluoromethyl-furan-2-yl)-6-(3-fluoro-pyridin-2-ylmethyl)-thieno[2,3-d]pyrimidin-4-yl]-bis-carbamicacid tert-butyl ester

A THF solution (1.5 mL) of acetic acid[4-tert-butoxycarbonylamino-2-(5-difluoromethyl-furan-2-yl)-thieno[2,3-d]pyrimidin-6-yl]-(3-fluoro-pyridin-2-yl)-methyl ester (100 mg, 0.16mmol) was purged with N₂ for ˜6 min, t-BuOH (23 μL, 0.24 mmol) was addedfollowed by a 0.1 M THF solution of SmI₂ (6.4 mL, 0.64 mmol). Afterstirring of the blue mixture for 25 min, the blue color changed toyellow and an additional 6.4 mL of the 0.1 M THF solution of SmI₂ wasadded. After 1 h saturated aqueous NH₄Cl was added and the organic layerwas separated. The aqueous layer was extracted with EtOAc and thecombined organics were dried (Na₂SO₄), concentrated, and purified bycolumn chromatography to yield 58 mg of the title compound.

Example 24 Step c2-(5-Difluoromethyl-furan-2-yl)-6-(3-fluoro-pyridin-2-ylmethyl)-thieno[2,3-d]pyrimidin-4-ylamine

Neat TFA (0.3 mL) was added to a CH₂Cl₂ solution (1.2 mL) of[2-(5-Difluoromethyl-furan-2-yl)-6-(3-fluoro-pyridin-2-ylmethyl)-thieno[2,3-d]pyrimidin-4-yl]-bis-carbamic acid tert-butyl ester (58 mg, 0.10mmol). After 30 min the mixture was concentrated in vacuo and theresidue was partitioned between saturated aqueous NaHCO₃ and EtOAc. Thelayers were separated and the organic layer was dried (Na₂SO₄) andconcentrated to give 37 mg of the title compound. ¹H NMR (300 MHz,Acetone-d₆) δ=8.41 (d, J=4.9 Hz, 1H), 7.62 (t, J=9.2 Hz, 1H), 7.40 (dt,J=4.4, 8.5 Hz, 1H), 7.33 (s, 1H), 7.14-7.23 (m, 1H), 7.00 (t, J=53.9 Hz,1H), 6.87-6.98 (m, 3H), 4.45 (d, J=1.9 Hz, 2H); MS m/e 377 (M+H).

Example 25[4-Amino-2-(5-difluoromethyl-furan-2-yl)-thieno[2,3-d]pyrimidin-6-yl]-(3-fluoro-pyridin-2-yl)-methanoneExample 25 Step a[2-(5-Difluoromethyl-furan-2-yl)-6-(3-fluoro-pyridine-2-carbonyl)-thieno[2,3-d]pyrimidin-4-yl]-bis-carbamicacid tert-butyl ester

Solid Dess-Martin periodinane (90 mg, 0.21 mmol) was added to a CH₂Cl₂solution (1.5 mL) of{2-(5-difluoromethyl-furan-2-yl)-6-[(3-fluoro-pyridin-2-yl)-hydroxy-methyl]-thieno[2,3-d]pyrimidin-4-yl}-bis-carbamicacid tert-butyl ester (94 mg, 0.16 mmol, an intermediate prepared inExample 23). After 1 h and saturated aqueous NaHCO₃ and saturatedaqueous Na₂S₂O₃ were added. The mixture was stirred vigorously for ˜15min and the aqueous phase was extracted with CH₂Cl₂. The combinedextracts were dried (Na₂SO₄), concentrated, and purified via columnchromatography to yield 71 mg of the title compound.

Example 25 Step b[4-Amino-2-(5-difluoromethyl-furan-2-yl)-thieno[2,3-d]pyrimidin-6-yl]-(3-fluoro-pyridin-2-yl)-methanone

Neat TFA (0.2 mL) was added to a CH₂Cl₂ solution (0.8 mL) of[2-(5-difluoromethyl-furan-2-yl)-6-(3-fluoro-pyridine-2-carbonyl)-thieno[2,3-d]pyrimidin-4-yl]-bis-carbamic acid tert-butyl ester (25 mg, 0.04mmol). After 1 h the mixture was concentrated in vacuo and the residuewas partitioned between saturated aqueous NaHCO₃ and EtOAc. The layerswere separated and the organic layer was dried (Na₂SO₄) and concentratedto give 16 mg of the title compound. ¹H NMR (300 MHz, DMSO-d₆) δ=8.63(d, J=4.5 Hz, 1H), 8.46 (s, 1H), 8.20 (br. s., 2H), 8.01 (t, J=9.6 Hz,1H), 7.81 (m, 1H), 7.30 (d, J=3.4 Hz, 1H), 7.17 (t, J=52.9 Hz, 1H),7.06-7.04 (m, 1H); MS m/e 391 (M+H).

Example 26[4-Amino-2-(5-difluoromethyl-furan-2-yl)-thieno[2,3-d]pyrimidin-6-yl]-(3-chloro-pyridin-2-yl)-methanol

The title compound was prepared using 3-chloro-pyridine-2-carbaldehydein place of 3-fluoro-pyridine-2-carbaldehyde as described in example 23.¹H NMR (300 MHz, Acetone-d₆) δ=8.63 (d, J=4.5 Hz, 1H), 7.95 (d, J=8.3Hz, 1H), 7.48 (dd, J=4.7, 7.7 Hz, 1H), 7.40 (s, 1H), 7.15-7.25 (m, 1H),6.91-7.12 (m, 3H), 7.00 (t, J=53.7 Hz, 1H), 6.37 (d, J=6.8 Hz, 1H),5.51-5.74 (m, 1H); MS m/e 409/411 (M+H).

Example 276-(3-Chloro-pyridin-2-ylmethyl)-2-(5-difluoromethyl-furan-2-yl)-thieno[2,3-d]pyrimidin-4-ylamine

[6-[(3-Chloro-pyridin-2-yl)-hydroxy-methyl]-2-(5-difluoromethyl-furan-2-yl)-thieno[2,3-d]pyrimidin-4-yl]-bis-carbamic acid tert-butyl ester (36 mg, 0.06 mmol,an intermediate prepared in Example 26), Et₃SiH (0.3 mL), and TFA (0.4mL) were combined and heated at 80° C. in the microwave for 1.5 h. Themixture was concentrated in vacuo, and the residue was partitionedbetween EtOAc and saturated aqueous NaHCO₃. The organic phase was dried(Na₂SO₄), concentrated, and purified by preparative thin layerchromatography to yield 15 mg of the title compound. ¹H NMR (300 MHz,Acetone-d₆) δ=8.46-8.58 (m, 1H), 7.88 (dd, J=1.5, 7.9 Hz, 1H), 7.36 (dd,J=4.7, 8.1 Hz, 1H), 7.32 (s, 1H), 7.14-7.22 (m, 1H), 7.00 (t, J=53.9 Hz,1H), 6.86-6.98 (m, 3H), 4.54 (s, 2H); MS m/e 393/395 (M+H).

Example 282-(2-Methoxy-pyridin-4-yl)-6-pyridin-2-ylmethyl-thieno[2,3-d]pyrimidin-4-ylamine

The title compound was prepared using[6-(hydroxy-phenyl-methyl)-2-(2-methoxy-pyridin-4-yl)-thieno[2,3-d]pyrimidin-4-yl]-bis-carbamicacid tert-butyl ester (an intermediate prepared in Example 22) in placeof[6-[(3-chloro-pyridin-2-yl)-hydroxy-methyl]-2-(5-difluoromethyl-furan-2-yl)-thieno[2,3-d]pyrimidin-4-yl]-bis-carbamicacid tert-butyl ester as described in Example 27. ¹H NMR (300 MHz,Acetone-d₆) δ=8.56 (d, J=4.5 Hz, 1H), 8.22 (d, J=5.3 Hz, 1H), 7.87 (d,J=5.7 Hz, 1H), 7.76 (m, 1H), 7.69 (s, 1H), 7.41 (d, J=7.9 Hz, 1H), 7.35(s, 1H), 7.21-7.31 (m, 1H), 6.93 (br. s., 2H), 4.40 (s, 2H), 3.93 (s,3H); MS m/e 350 (M+H).

Example 29[4-Amino-2-(5-difluoromethyl-furan-2-yl)-thieno[2,3-d]pyrimidin-6-yl]-(3-methoxy-pyridin-2-yl)-methanol

The title compound was prepared using 3-methoxy-pyridine-2-carbaldehydein place of 3-fluoro-pyridine-2-carbaldehyde as described in example 23.¹H NMR (300 MHz, Acetone-d₆) δ=8.20 (d, J=4.1 Hz, 1H), 7.51 (d, J=8.3Hz, 1H), 7.33-7.46 (m, 2H), 7.08-7.22 (m, 1H), 6.99 (t, J=53.5 Hz, 1H),6.85-6.98 (m, 2H), 6.22 (s, 1H), 3.92 (s, 3H); MS m/e 405 (M+H).

Example 301-[4-Amino-2-(5-difluoromethyl-furan-2-yl)-thieno[2,3-d]pyrimidin-6-yl]-1-(3-fluoro-pyridin-2-yl)-ethanol

A 3.0 M THF solution of MeMgCl (0.04 mL, 0.12 mmol) was added to a 4° C.THF solution (1 mL) of[2-(5-difluoromethyl-furan-2-yl)-6-(3-fluoro-pyridine-2-carbonyl)-thieno[2,3-d]pyrimidin-4-yl]-bis-carbamicacid tert-butyl ester (44 mg, 0.07 mmol, an intermediate prepared inExample 25). After 30 min at 4° C. the mixture was warmed to rt. After 1h at rt an additional 0.04 mL of the 3.0 M THF solution of MeMgCl wasadded and the mixture was stirred overnight. Saturated aqueous NH₄Cl wasadded and the aqueous phase was extracted with CH₂Cl₂. The combinedorganics were dried (Na₂SO₄), concentrated, and purified by columnchromatography to yield 25 mg of white solid that was used withoutfurther purification. A solution of this solid in 0.4 mL of CH₂Cl₂ wastreated with neat TFA (0.1 mL). After 1 h The mixture was concentratedin vacuo, and the residue was partitioned between EtOAc and saturatedaqueous NaHCO₃. The organic phase was dried (Na₂SO₄) and concentrated toyield 19 mg of the title compound that was analytically pure. ¹H NMR(300 MHz, Acetone-d₆) δ=8.48 (d, J=4.1 Hz, 1H), 7.63-7.76 (m, 1H),7.52-7.61 (m, 1H), 7.50 (s, 1H), 7.19 (d, J=3.4 Hz, 1H), 7.00 (t, J=53.9Hz, 1H), 6.91-7.00 (m, 3H), 6.46 (br. s., 1H), 2.05 m/e 407 (M+H).

Example 31[4-Amino-2-(5-difluoromethyl-furan-2-yl)-thieno[2,3-d]pyrimidin-6-yl]-pyridin-2-yl-methanol

The title compound was prepared using pyridine-2-carbaldehyde in placeof 3-fluoro-pyridine-2-carbaldehyde as described in Example 23. ¹H NMR(300 MHz, Acetone-d₆) δ=8.56 (d, J=3.8 Hz, 1H), 7.75-7.90 (m, 1H), 7.65(d, J=7.9 Hz, 1H), 7.46 (s, 1H), 7.32 (dd, J=4.9, 7.5 Hz, 1H), 7.10-7.20(m, 1H), 7.00 (t, J=53.5 Hz, 1H), 6.90-7.00 (m, 3H), 6.11 (s, 1H), 5.87(br. s., 1H); MS m/e 375 (M+H).

Example 32[4-Amino-2-(5-difluoromethyl-furan-2-yl)-thieno[2,3-d]pyrimidin-6-yl]-(3-bromo-pyridin-2-yl)-methanol

The title compound was prepared using 3-bromo-pyridine-2-carbaldehyde inplace of 3-fluoro-pyridine-2-carbaldehyde as described in Example 23. ¹HNMR (300 MHz, MeOD-d₄) δ=8.81 (m, 1H), 8.52-8.72 (m, 1H), 7.63-7.92 (m,2H), 7.46-7.63 (m, 1H), 7.05 (m, 1H), 6.92 (t, J=53.7 Hz, 1H), 6.63 (br.s., 1H); MS m/e 453/455 (M+H).

Example 336-(3-Bromo-pyridin-2-ylmethyl)-2-(5-difluoromethyl-furan-2-yl)-thieno[2,3-d]pyrimidin-4-ylamine

The title compound was prepared using[6-[(3-bromo-pyridin-2-yl)-hydroxy-methyl]-2-(5-difluoromethyl-furan-2-yl)-thieno[2,3-d]pyrimidin-4-yl]-bis-carbamicacid tert-butyl ester (an intermediate prepared in Example 32) in placeof{2-(5-difluoromethyl-furan-2-yl)-6-[(3-fluoro-pyridin-2-yl)-hydroxy-methyl]-thieno[2,3-d]pyrimidin-4-yl}-bis-carbamicacid tert-butyl ester as described in Example 24. ¹H NMR (300 MHz,Acetone-d₆) δ=8.50-8.61 (m, 1H), 7.97-8.08 (m, 1H), 7.31-7.33 (m, 1H),7.27 (dd, J=4.7, 8.1 Hz, 1H), 7.11-7.21 (m, 1H), 7.00 (t, J=53.9 Hz,1H), 6.92-6.97 (m, 3H), 4.56 (s, 2H); MS m/e 437/439 (M+H).

Example 342-{[4-Amino-2-(5-difluoromethyl-furan-2-yl)-thieno[2,3-d]pyrimidin-6-yl]-hydroxy-methyl}-pyridin-3-ol

The title compound was prepared using 3-hydroxy-pyridine-2-carbaldehydein place of 3-fluoro-pyridine-2-carbaldehyde as described in example 23.¹H NMR (300 MHz, Acetone-d₆) δ=8.09-8.17 (m, 1H), 7.41 (s, 1H),7.21-7.36 (m, 2H), 7.18 (d, J=3.4 Hz, 1H), 6.99 (t, J=53.9 Hz, 1H),6.87-6.98 (m, 2H), 6.25 (s, 1H); MS m/e 391 (M+H).

Example 352-(5-Difluoromethyl-furan-2-yl)-6-pyridin-2-ylmethyl-thieno[2,3-d]pyrimidin-4-ylamine

The title compound was prepared using[2-(5-difluoromethyl-furan-2-yl)-6-(hydroxy-pyridin-2-yl-methyl)-thieno[2,3-d]pyrimidin-4-yl]-bis-carbamicacid tert-butyl ester (an intermediate prepared in Example 31) in placeof{2-(5-difluoromethyl-furan-2-yl)-6-[(3-fluoro-pyridin-2-yl)-hydroxy-methyl]-thieno[2,3-d]pyrimidin-4-yl}-bis-carbamicacid tert-butyl ester as described in Example 24. ¹H NMR (300 MHz,MeOD-d₄) δ=8.86 (d, J=5.7 Hz, 1H), 8.54-8.69 (m, 1H), 7.96-8.13 (m, 2H),7.53-7.68 (m, 2H), 7.04-7.13 (m, 1H), 6.97 (t, J=53.6 Hz, 1H), 4.84 (s,2H); MS m/e 359 (M+H).

Example 361-[4-Amino-2-(5-difluoromethyl-furan-2-yl)-thieno[2,3-d]pyrimidin-6-yl]-1-pyridin-2-yl-ethanolExample 36 Step a[2-(5-Difluoromethyl-furan-2-yl)-6-(pyridine-2-carbonyl)-thieno[2,3-d]pyrimidin-4-yl]-bis-carbamicacid tert-butyl ester

The title compound was prepared using[2-(5-difluoromethyl-furan-2-yl)-6-(hydroxy-pyridin-2-yl-methyl)-thieno[2,3-d]pyrimidin-4-yl]-bis-carbamicacid tert-butyl ester (an intermediate prepared in Example 31) in placeof{2-(5-difluoromethyl-furan-2-yl)-6-[(3-fluoro-pyridin-2-yl)-hydroxy-methyl]-thieno[2,3-d]pyrimidin-4-yl}-bis-carbamicacid tert-butyl ester as described in Example 25.

Example 36 Step b1-[4-Amino-2-(5-difluoromethyl-furan-2-yl)-thieno[2,3-d]pyrimidin-6-yl]-1-pyridin-2-yl-ethanol

The title compound was prepared using[2-(5-difluoromethyl-furan-2-yl)-6-(pyridine-2-carbonyl)-thieno[2,3-d]pyrimidin-4-yl]-bis-carbamicacid tert-butyl ester in place of [2-(5-difluoromethyl-furan-2-yl)-6-(3-fluoro-pyridine-2-carbonyl)-thieno[2,3 -d]pyrimidin-4-yl]-bis-carbamicacid tert-butyl ester as described in Example 30. ¹H NMR (300 MHz,Acetone-d₆) δ=8.52 (d, J=4.5 Hz, 1H), 7.79 (t, J=7.5 Hz, 1H), 7.71 (d,J=7.9 Hz, 1H), 7.48 (s, 1H), 7.27 (t, J=6.0 Hz, 1H), 7.15 (d, J=3.4 Hz,1H), 6.96 (t, J=53.5 Hz, 1H), 6.90-6.96 (m, 3H), 1.99 (s, 3H); MS m/e389 (M+H).

Example 372-(5-Difluoromethyl-furan-2-yl)-6-(3-methoxy-pyridin-2-ylmethyl)-thieno[2,3-d]pyrimidin-4-ylamine

The title compound was prepared using{2-(5-difluoromethyl-furan-2-yl)-6-[hydroxy-(3-methoxy-pyridin-2-yl)-methyl]-thieno[2,3-d]pyrimidin-4-yl}-bis-carbamicacid tert-butyl ester (an intermediate prepared in Example 29) in placeof{2-(5-difluoromethyl-furan-2-yl)-6-[(3-fluoro-pyridin-2-yl)-hydroxy-methyl]-thieno[2,3-d]pyrimidin-4-yl}-bis-carbamicacid tert-butyl ester as described in Example 24. ¹H NMR (300 MHz,MeOD-d₄) δ=8.42 (d, J=4.5 Hz, 1H), 8.31-8.34 (m, 1H), 7.95-8.12 (m, 1H),7.50-7.70 (m, 2H), 7.05-7.13 (m, 1H), 6.97 (t, J=53.3 Hz, 1H), 4.74 (s,2H), 4.16 (s, 3H); MS m/e 389 (M+H)

Biological Assays and Activity

Ligand Binding Assay for Adenosine A2a Receptor (A2A-B)

Ligand binding assay of adenosine A2a receptor was performed usingplasma membrane of HEK293 cells containing human A2a adenosine receptor(PerkinElmer, RB-HA2a) and radioligand [³H]CGS21680 (PerkinElmer,NET1021). Assay was set up in 96-well polypropylene plate in totalvolume of 200 μL by sequentially adding 20 μL 1:20 diluted membrane, 130μL assay buffer (50 mM Tris.HCl, pH7.4 10 mM MgCl₂, 1 mM EDTA)containing [³H] CGS21680, 50 μL diluted compound (4×) or vehicle controlin assay buffer. Nonspecific binding was determined by 80 mM NECA.Reaction was carried out at room temperature for 2 hours beforefiltering through 96-well GF/C filter plate pre-soaked in 50 mMTris.HCl, pH7.4 containing 0.3% polyethylenimine. Plates were thenwashed 5 times with cold 50 mM Tris.HCl, pH7.4, dried and sealed at thebottom. Microscintillation fluid 30 μL was added to each well and thetop sealed. Plates were counted on Packard Topcount for [³H]. Data wasanalyzed in Microsoft Excel and GraphPad Prism programs. (Varani, K.;Gessi, S.; Dalpiaz, A.; Borea, P. A. British Journal of Pharmacology,1996, 117, 1693)

Adenosine A2a Receptor Functional Assay (A2AGAL2)

To initiate the functional assay, cryopreserved CHO-K1 cellsoverexpressing the human adenosine A2a receptor and containing a cAMPinducible beta-galactosidase reporter gene were thawed, centrifuged,DMSO containing media removed, and then seeded with fresh culture mediainto clear 384-well tissue culture treated plates (BD #353961) at aconcentration of 10K cells/well. Prior to assay, these plates werecultured for two days at 37° C., 5% CO₂, 90% Rh. On the day of thefunctional assay, culture media was removed and replaced with 45 uLassay medium (Hams/F-12 Modified (Mediatech #10-080CV) supplementedw/0.1% BSA). Test compounds were diluted and 11 point curves created ata 1000× concentration in 100% DMSO. Immediately after addition of assaymedia to the cell plates, 50 nL of the appropriate test compoundantagonist or agonist control curves were added to cell plates using aCartesian Hummingbird. Compound curves were allowed to incubate at roomtemperature on cell plates for approximately 15 minutes before additionof a 15 nM NECA (Sigma E2387) agonist challenge (5 uL volume). A controlcurve of NECA, a DMSO/Media control, and a single dose of Forskolin(Sigma F3917) were also included on each plate. After additions, cellplates were allowed to incubate at 37° C., 5% CO₂, 90% Rh for 5.5-6hours. After incubation, media was removed, and cell plates were washed1×50 uL with DPBS w/o Ca & Mg (Mediatech 21-031-CV). Into dry wells, 20uL of 1× Reporter Lysis Buffer (Promega E3971 (diluted in dH₂O from 5×stock)) was added to each well and plates frozen at −20° C. overnight.For β-galactosidase enzyme colorimetric assay, plates were thawed out atroom temperature and 20μL 2× assay buffer (Promega) was added to eachwell. Color was allowed to develop at 37° C., 5% CO₂, 90% Rh for 1-1.5 hor until reasonable signal appeared. The calorimetric reaction wasstopped with the addition of 60 μL/well 1M sodium carbonate. Plates werecounted at 405 nm on a SpectraMax Microplate Reader (Molecular Devices).Data was analyzed in Microsoft Excel and IC/EC50 curves were fit using astandardized macro.

Adenosine A1 Receptor Functional Assay (A1GAL2)

To initiate the functional assay, cryopreserved CHO-K1 cellsoverexpressing the human adenosine A1 receptor and containing a cAMPinducible beta-galactosidase reporter gene were thawed, centrifuged,DMSO containing media removed, and then seeded with fresh culture mediainto clear 384-well tissue culture treated plates (BD #353961) at aconcentration of 10K cells/well. Prior to assay, these plates werecultured for two days at 37° C., 5% CO₂, 90% Rh. On the day of thefunctional assay, culture media was removed and replaced with 45 uLassay medium (Hams/F-12 Modified (Mediatech #10-080CV) supplementedw/0.1% BSA). Test compounds were diluted and 11 point curves created ata 1000× concentration in 100% DMSO. Immediately after addition of assaymedia to the cell plates, 50 nL of the appropriate test compoundantagonist or agonist control curves were added to cell plates using aCartesian Hummingbird. Compound curves were allowed to incubate at roomtemperature on cell plates for approximately 15 minutes before additionof a 4 nM r-PIA (Sigma P4532)/1 uM Forskolin (Sigma F3917) agonistchallenge (5 uL volume). A control curve of r-PIA in 1 uM Forskolin, aDMSO/Media control, and a single dose of Forskolin were also included oneach plate. After additions, cell plates were allowed to incubate at 37°C., 5% CO₂, 90% Rh for 5.5-6 hours. After incubation, media was removed,and cell plates were washed 1×50 uL with DPBS w/o Ca & Mg (Mediatech21-031-CV). Into dry wells, 20 uL of 1× Reporter Lysis Buffer (PromegaE3971 (diluted in dH₂O from 5× stock)) was added to each well and platesfrozen at −20° C. overnight. For β-galactosidase enzyme colorimetricassay, plates were thawed out at room temperature and 20 μL 2× assaybuffer (Promega) was added to each well. Color was allowed to develop at37° C., 5% CO₂, 90% Rh for 1-1.5 h or until reasonable signal appeared.The colorimetric reaction was stopped with the addition of 60 μL/well 1Msodium carbonate. Plates were counted at 405 nm on a SpectraMaxMicroplate Reader (Molecular Devices). Data was analyzed in MicrosoftExcel and IC/EC50 curves were fit using a standardized macro.

A2a ASSAY DATA Example A2AGAL2 Ki μM A2A-B Ki μM A1GAL2 Ki μM 1 0.126212ND 1.58708 2 ND ND ND 3 0.0066819 ND 0.352777 4 ND ND ND 5 ND ND ND 6 NDND ND 7 ND ND ND 8 ND ND ND 9 ND ND ND 10 ND ND ND 11 ND ND ND 12 ND NDND 13 ND ND ND 14 ND ND ND 15 0.00966273 ND 0.172147 16 0.00118005 ND0.0299364 17 0.00709414 ND 0.100462 18 0.0300124 ND 0.489441 19 0.409449ND 0.579162 20 0.129867 ND 0.33721 21 0.00525049 ND 0.0281838 22 ND NDND 23 0.00377051 ND 0.0595388 24 0.00620869 ND 0.092619 25 0.0248599 ND0.838301 26 ND ND ND 27 ND ND ND 28 ND ND ND 29 0.00239166 ND 0.024877130 0.0113684 0.0162817 0.40281 31 0.00481947 ND 0.0469029 32 0.0016248ND 0.0235342 33 0.00217721 ND 0.0212031 34 0.00428845 ND 0.110586 350.00473151 ND 0.0618871 36 0.0359832 ND 0.267055 37 0.00228402 ND0.0326137 ND indicates that no data was available.

While the foregoing specification teaches the principles of the presentinvention, with examples provided for the purpose of illustration, itwill be understood that the practice of the invention encompasses all ofthe usual variations, adaptations and/or modifications as come withinthe scope of the following claims and their equivalents.

All publications disclosed in the above specification are herebyincorporated by reference in full.

1. A compound of Formula Z

wherein X is selected from the group consisting of

R¹ is heteroaryl which may be substituted with one substituent selectedfrom the group consisting of Cl, Br, F, OH, CN, C₍₁₋₄₎alkyl, CHF₂, CF₃,OCF₃, cyclopropyl, and OC₍₁₋₄₎alkyl; R² is heteroaryl wherein saidheteroaryl is optionally substituted with Cl, F, Br, OC₍₁₋₄₎alkyl, OCF₃,OH, C₍₁₋₄₎alkyl, CHF₂, CF₃, OCH₂CF₃, or a ring selected from the groupconsisting of:

wherein R^(a), R^(b), and R^(c) are independently H or C₍₁₋₄₎alkyl;R^(d) is H, —C₍₁₋₄₎alkyl, —CH₂CH₂OCH₂CH₂OCH₃, —CH₂CO₂H,—C(O)C₍₁₋₄₎alkyl, or —CH₂C(O)C₍₁₋₄₎alkyl; and solvates, hydrates,tautomers and pharmaceutically acceptable salts thereof.
 2. A compoundof claim 1, wherein: R¹ is furyl, oxazolyl, thiazolyl, pyridyl,pyrimadyl, isoxazolyl, pyrrolyl, imidazoyl, or pyridazyl, any of whichmay be substituted with one substituent selected from the groupconsisting of Cl, Br, F, OH, CN, C₍₁₋₄₎alkyl, CHF₂, CF₃, cyclopropyl,and OC₍₁₋₄₎alkyl; R² is pyrimadyl, isoxazolyl, pyrrolyl, imidazoyl,furyl, oxazolyl, pyridyl or pyridazyl, any of which may be substitutedwith one substituent selected from the group consisting of Cl, Br, F,OH, C₁₋₄₎alkyl, CHF₂, CF₃, and OC₍₁₋₄₎alkyl; and solvates, hydrates,tautomers and pharmaceutically acceptable salts thereof.
 3. A compoundof claim 2, wherein: X is selected from the group consisting of

R¹ is furyl, oxazolyl, thiazolyl, pyridyl, or pyridazyl, any of whichmay be substituted with one substituent selected from the groupconsisting of Cl, Br, F, OH, CN, C₍₁₋₄₎alkyl, CHF₂, CF₃, cyclopropyl,and OC₍₁₋₄₎alkyl; R² is pyridyl or pyridazyl, either of which may besubstituted with one substituent selected from the group consisting ofCl, Br, F, OH, C₍₁₋₄₎alkyl, CHF₂, CF₃, and OC₁₋₄₎alkyl; and solvates,hydrates, tautomers and pharmaceutically acceptable salts thereof.
 4. Acompound of claim 3, wherein: R¹ is furyl, oxazolyl, thiazolyl, pyridyl,or pyridazyl, any of which may be substituted with one substituentselected from the group consisting of CN, CH₃, CHF₂, cyclopropyl, andOCH₃; R² is pyridyl or pyridazyl, either of which may be substitutedwith one substituent selected from the group consisting of Cl, Br, F,OH, and OCH₃; and solvates, hydrates, tautomers and pharmaceuticallyacceptable salts thereof.
 5. A compound of claim 4, wherein: R¹ isselected from the group consisting of:

R² is selected from the group consisting of:

and solvates, hydrates, tautomers and pharmaceutically acceptable saltsthereof.
 6. A compound selected from the group consisting of:

and solvates, hydrates, tautomers, and pharmaceutically acceptable saltsthereof
 7. A pharmaceutical composition comprising the compound of claim1; and a pharmaceutically acceptable carrier.
 8. A method of treating asubject having a disorder ameliorated by antagonizing Adenosine A2areceptors in appropriate cells in the subject, which comprisesadministering to the subject a therapeutically effective dose of thecompound of claim
 1. 9. A method of preventing a disorder ameliorated byantagonizing Adenosine A2a receptors in appropriate cells in thesubject, comprising administering to the subject a prophylacticallyeffective dose of the compound of claim 1 either preceding or subsequentto an event anticipated to cause a disorder ameliorated by antagonizingAdenosine A2a receptors in appropriate cells in the subject.
 10. Themethod of treating a subject having a disorder ameliorated byantagonizing Adenosine A2a receptors in appropriate cells in thesubject, comprising administering to the subject a therapeutically orprophylactically effective dose of the pharmaceutical composition ofclaim
 7. 11. The method of preventing a disorder ameliorated byantagonizing Adenosine A2a receptors in appropriate cells in thesubject, comprising administering to the subject a therapeutically orprophylactically effective dose of the pharmaceutical composition ofclaim
 7. 12. The method of claim 8, wherein the disorder is aneurodegenerative disorder or a movement disorder.
 13. The method ofclaim 8, wherein the disorder is selected from the group consisting ofParkinson's Disease, Huntington's Disease, Multiple System Atrophy,Corticobasal Degeneration, Alzheimer's Disease, and Senile Dementia. 14.The method of claim 9, wherein the disorder is a neurodegenerativedisorder or a movement disorder.
 15. The method of claim 9, wherein thedisorder is selected from the group consisting of Parkinson's Disease,Huntington's Disease, Multiple System Atrophy, CorticobasalDegeneration, Alzheimer's Disease, and Senile Dementia.
 16. The methodof claim 8, wherein the disorder is Parkinson's Disease.
 17. The methodof claim 8, wherein the disorder is addiction.
 18. The method of claim8, wherein the disorder is Attention Deficit Hyperactivity Disorder(ADHD).
 19. The method of claim 8, wherein the disorder is depression.20. The method of claim 8, wherein the disorder is anxiety.